OBJECTIVE： To study the anti－ metastatic action of equal/unequal－ fork－ peptide of YIGSR.METHODS： To observe the influence of drugs on metastasis of melanoma B16 to lung in mice.RESULTS： The metastatic rate of lung in 46 mice of control and experimental groups was 100％ ,21 days after inoculating melanoma B16 cells via caudal vein.Injection of YIGSR in combination with tumor cells could reduce the number of metastatic nodules in a dose－ dependent manner.The numbers of metastatic nodules in 100μ g～ 200μ g equal fork peptide group and same dosage unequal fork peptide group were significantly different from that in control group(P<0.05;P<0.01 respectively).50μ g equal fork peptide group was different from control group.The synthetic peptide could not decrease the weight of the lung with metastatic lesions.CONCLUSION： YIGSR derivants have effective antimetastatic actions.Its mechanism may related to the inhibiting effect on formation of cancer cell emboli,which retain in microvessels of remote target organs,and multiplicate and penetrate blood vessels to form micrometastatic lesions.

M2 protein of influenza A virus is encoded by a spliced mRNA derived from RNA segment 7 and plays an important role in influenza virus replication. It is also a target molecule of anti-virus drugs. We extracted the viral genome RNAs from MDCK cells infected with swine influenza A virus (SIV) H3N2 subtype and amplified the SIV M2 gene by reverse transcriptase-polymerase chain reaction using the isloated viral genome RNAs as template. The amplified cDNA was cloned into a prokaryotic expression vector pET-28a(+) (designated pET-28a(+)-M2) and a eukaryotic expression vector p3xFLAG-CMV-7.1 (designated p3xFLAG-CMV-7.1-M2), respectively. The resulted constructs were confirmed by restriction enzyme digestion and DNA sequencing analysis. We then transformed the plasmid pET-28a(+)-M2 into Escherichia coli BL21 (DE3) strain and expressed it by adding 1 mmol/L of IPTG (isopropyl-beta-D-thiogalactopyranoside). The recombinant M2 protein was purified from the induced bacterial cells using Ni(2+) affinity chromatography. Wistar rats were immunized with the purified M2 protein for producing polyclonal antibodies specific for it. Western blotting analysis and immunofluorescence analysis showed that the produced antibodies were capable of reacting with M2 protein expressed in p3xFLAG-CMV-7.1-M2-transfected cells as well as that synthesized in SIV-infected cells. We also transfected plasmid p3xFLAG-CMV-7.1-M2 into Vero cells and analyzed its subcellular localization by immunofluorescence. The M2 protein expressed in the Vero cells was 20 kDa in size and dominantly localized in the cytoplasm, showing a similar distribution to that in SIV-infected cells. Western blotting analysis of SIV-infected cells suggested that M2 was a late phase protein, which was detectable 12 h post-infection, later than NS1, NP and M1 proteins. It would be a potential molecular indicator of late phases replication of virus. Our results would be useful for studying the biological function of M2 protein in SIV replication.
Animals ; Antibodies, Monoclonal ; biosynthesis ; Cercopithecus aethiops ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Influenza A Virus, H3N2 Subtype ; genetics ; RNA ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Swine ; Transfection ; Vero Cells ; Viral Matrix Proteins ; biosynthesis ; genetics ; Virus Replication ; genetics

Abstract: The new generation of artificial intelligence technology, represented by deep learning, has emerged as a crucial driving force in the advancement of new drug research and development. This article creatively proposes a workflow named “Molecular Factory” for the design and optimization of drug molecules based on artificial intelligence technology. This workflow integrates intelligent molecular generation models, high-performance molecular docking algorithms, and accurate protein-ligand binding affinity prediction methods. It has been integrated as a core module into DrugFlow, a one-stop drug design software platform, providing a comprehensive set of mature solutions for the discovery and optimization of lead compounds. Utilizing the “Molecular Factory” module, we conducted the research of second-generation inhibitors against Menin that can combat drug resistance. Through the integration of computational and experimental approaches, we rapidly identified multiple promising compounds. Among them, compound RG-10 exhibited the IC50 values of 9.681 nmol/L, 233.2 nmol/L, and 40.09 nmol/L against the wild-type Menin, M327I mutant, and T349M mutant, respectively. Compared to the positive reference molecule SNDX-5613, which has entered Phase II clinical trials, RG-10 demonstrated significantly enhanced inhibitory activity against the M327I and T349M mutants. These findings fully demonstrate the unique advantages of the "Molecular Factory" technology in practical drug design and development scenarios. It can rapidly and efficiently generate high-quality active molecules targeting specific protein structures, holding significant value and profound implications for advancing new drug discovery.

Objective To analyze the changes in follicular helper T (Tfh) cells during HIV-1 in-fection, to investigate the influences of Tfh cells and Tfh-related molecules on HIV-1 progression and to pro-vide references for further research on using Tfh cells in highly active antiretroviral therapy ( HAART) and vaccines. Methods This study enrolled 33 patients with HIV-1 infection, including 11 long-term nonpro-gressors (LTNP), 10 rapid progressors (RP) and 12 typical progressors (TP), and 11 healthy subjects (normal controls, NC). Peripheral blood mononuclear cells were isolated from each subject. Multicolor flow cytometry was performed to detect CD4+CD45RA-CXCR5+Tfh and CD4+CD45RA-CXCR3-CXCR5+PD-1+Tfh subsets and the levels of inducible costimulatory molecule (ICOS), IFN-γ and IL-21. Moreover, the levels of IL-10 and the percentages of CD19+B cells in plasma samples of each group were also analyzed. Relationships among Tfh, CD4 and B cells were analyzed. Results The percentages of both Tfh subsets were higher in patients with HIV-1 infection than in NC. Compared with NC, LTNP had the highest percent-age of CD4+CD45RA-CXCR3-CXCR5+PD-1+Tfh cells (P<0. 05). Expression of Tfh-related molecules ICOS, IFN-γ and IL-21 were enhanced significantly upon Staphylococcus enterotoxin B ( SEB) stimulation, ICOS+Tfh cells were negatively related with HIV-1 progression, but had a positive correlation with CD19+B cells (r=-0. 49, P<0. 01; r=0. 60, P<0. 05). IL-10 level in plasma increased significantly in patients withHIV-1 infection , especially in TP and RP ( TP vs NC : P＜0. 01 ; RP vs NC : P<0. 05 ) . Conclusion HIV-1 patients and NC had significant differences in the expression of Tfh cells and Tfh-related molecules in peripheral blood. ICOS+Tfh cells were closely related to the progression of HIV-1 infection and the function of B cells.